Search Results (2,051)

Background: Polymicrobial skin and soft tissue infections (SSTIs) are frequently complicated by methicillin-resistant Staphylococcus aureus (MRSA) and co-colonizing Gram-negative pathogens like Pseudomonas aeruginosa (P. aeruginosa). Mupirocin, the clinical gold standard, is limited by rising resistance and an intrinsic “mupirocin gap” against P. aeruginosa. This study evaluates a novel Nigella sativa (NS) seed oil topical formulation as an alternative. Methods: A 4-tier preclinical platform assessed the NS formulation against MRSA, methicillin-sensitive S. aureus (MSSA), Streptococcus pyogenes, and P. aeruginosa. The pipeline included: (1) in vitro agar diffusion, (2) a gauze biofilm prevention model, (3) an ex vivo porcine ear skin model challenging epidermal lipid barriers, and (4) an in vivo Galleria mellonella model evaluating trans-cuticular systemic protection. Results: The NS formulation produced extensive diffusion zones, completely inhibiting S. pyogenes and outperforming controls against MSSA and P. aeruginosa. In the gauze model, NS achieved complete eradication of MSSA and S. pyogenes, while significantly suppressing MRSA and P. aeruginosa biofilms (p < 0.001). In the ex vivo porcine model, NS yielded >1.5 to >2.5 log reductions across all pathogens at 24 h (p < 0.001). Furthermore, in the in vivo G. mellonella model, topical NS significantly reduced the systemic bioburden of MSSA, S. pyogenes, and P. aeruginosa (p < 0.001), though MRSA reduction lacked statistical significance. Conclusions: The novel NS formulation demonstrates potent broad-spectrum antimicrobial activity. By effectively bridging the “mupirocin gap” against P. aeruginosa and demonstrating significant efficacy against MRSA in in vitro and ex vivo environments, it represents a promising plant-based pre-clinical candidate that strongly warrants future evaluation in live mammalian wound healing models. Full article

Background/Objectives: Immune surveillance is increasingly recognized as a modifier of myeloproliferative neoplasm (MPN) initiation and evolution, yet the contribution of the NKG2D receptor and its ligands MICA/MICB to CALR-mutated disease remains unclear. Methods: We performed high-resolution next-generation sequencing genotyping of MICA and MICB in 43 patients with CALR-mutated MPN (WHO 2022 criteria) and compared the allele and haplotype distributions with those of 156 healthy Bulgarian controls and 85 patients with JAK2 V617F-positive MPN. Associations were tested using age- and sex-adjusted additive generalized linear models; bi-locus haplotypes were evaluated using haplotype score methods. In a genotyped subgroup (35 CALR-mutated MPN patients and 105 controls), functional KLRK1 (NKG2D) polymorphisms were analyzed for haplotype-level associations. We also performed 700 ns molecular dynamics simulations of selected MICA variants in complex with NKG2D and reanalyzed publicly available single-cell RNA-sequencing data (GSE117826) and RNA-sequencing data from CRISPR/Cas9-edited CALR-mutant iPSC-derived megakaryocytes to evaluate MICA/MICB expression. Results: MICA*004:001 was significantly associated with CALR-mutated MPN versus controls (p = 0.004; Bonferroni-adjusted p = 0.047), while MICB*008:001 showed only nominal association. Exploratory haplotype analyses identified a MICA*009:01-MICB*004:001 haplotype associated with CALR-mutated status (p = 0.008) and a KLRK1 G-A-G-T haplotype (rs1049174-rs2617160-rs2246809-rs2617170) associated with increased CALR-mutated MPN risk (OR = 3.61; p = 0.029). Transcriptomic reanalysis indicated a higher fraction of CALR-mutant stem and progenitor cells expressing detectable MICA/MICB transcripts, and heterozygous CALR-mutant megakaryocytes exhibited higher MICA expression than the wild type. Conclusions: Together, these data support an exploratory immunogenetic and transcriptomic link between the NKG2D-MICA/MICB axis and CALR-mutated MPN, but direct protein-level and functional studies are required before mechanistic or therapeutic conclusions can be drawn. Full article

This study focuses on the synthesis of glucovanillin mediated by UGT109A1 and its mechanism against Type 2 Diabetes Mellitus (T2DM). Recombinant UGT109A1 successfully synthesized glucovanillin from vanillin using UDP-Glc as the sugar donor. Through network pharmacology, 140 potential targets were identified. Seven key targets were further screened using LASSO and SVM-RFE algorithms. Among these, SLC5A1 and ADK showed strong diagnostic potential, with AUC values ranging from 0.85 to 0.89. Immune infiltration analysis linked these core targets to M2 macrophages. Single-cell transcriptomics revealed that ADK is widely expressed but enriched in B cells, while TLR9 is confined to plasmacytoid dendritic cells (pDCs). Cell-to-cell communication analysis identified a pDC-to-B cell signaling axis. In vitro assays demonstrated that glucovanillin exhibits concentration-dependent inhibitory activity against α-glucosidase with moderate potency, with an IC₅₀ of 413.84 ± 12.80 μM. Molecular docking, 200 ns molecular dynamics simulations (MD), and MM/PBSA calculations showed that glucovanillin binds more strongly to α-glucosidase (−7.4 kcal/mol) than vanillin (−5.4 kcal/mol). Therefore, the glycosylation mediated by UGT109A1 enhanced the bioactivity and targeting specificity of vanillin. In summary, glucovanillin exerts anti-T2DM effects through a dual mechanism involving α-glucosidase inhibition and regulation of key targets, making it a promising lead compound for T2DM treatment. Full article

Background and Aims: The “quantity” of bone can be evaluated by dual-energy X-ray absorptiometry (DXA) scans, but not its “quality. We aim to study the clinical relevance of urinary-N-terminal telopeptide (NTx) in a retrospective exploratory study. Patients and Methods: The medical records of patients with osteoporosis, osteopenia with or without fractures, and with available urinary NTx were retrospectively reviewed; those on anti-osteoporotic medication before the start of the study were excluded. In all NTx levels, bone-specific alkaline phosphatase (BSAP), parathormone, serum calcium, and vitamin D were measured. In all cases, a recent DXA scan and fracture risk assessment (FRAX) had been performed. Appropriate statistics were applied using SPSS. 15. Results: Included were 93 patients (17.2% males); thirty-one (33.33%) had osteoporosis, 56 (60.21%) osteopenia, whereas 36 (38.7%) had prior or existing fractures. Older participants had lower NTx levels, and females had higher NTx levels, albeit NS. A negative correlation was found between the T-score of the left hip and NTx levels (p = 0.015) but not of the right hip or lumbar spine. In multivariate analysis, NTx levels (p = 0.013) and FRAX (p = 0.001) were significantly associated with fractures. Patients with osteoporosis had higher NTx levels when compared to patients with osteopenia (p = 0.015). NTx at a cut-off value of 207.4 showed a sensitivity of 80.6% and a specificity of 56.1% for the diagnosis of previous fracture with an area under the curve (AUC) of 0.72 (95% CI: 0.61, 0.83). Conclusions: Elevated NTx levels were significantly associated with existing or prior fractures. Combining DXA scan and FRAX, with NTx testing, may provide a comprehensive approach to osteoporosis assessment and treatment. Further prospective studies are warranted to validate its clinical utility. Full article

Tourism destination marketers increasingly rely on video advertising, yet the psychological mechanisms linking perceived advertising design and destination familiarity to engagement remain underspecified in tourism contexts. Drawing on narrative transportation and advertising stimulation perspectives, this study examines how perceived advertising design and destination familiarity relate to narrative transportation and advertising stimulation, and how these mechanisms relate to engagement. Using a survey of 915 Portuguese respondents and structural equation modeling in AMOS, we estimate a model comprising advertising design, destination familiarity, narrative transportation, advertising stimulation, and engagement. Results show that perceived advertising design is positively associated with narrative transportation (β = 0.451, p < 0.01) and advertising stimulation (β = 0.158, p < 0.01). Destination familiarity is also positively associated with narrative transportation (β = 0.215, p < 0.01) and advertising stimulation (β = 0.104, p < 0.01). Narrative transportation is strongly associated with advertising stimulation (β = 0.659, p < 0.01), whereas narrative transportation does not show a significant direct association with engagement (β = 0.086, n.s.). Advertising stimulation is positively associated with engagement (β = 0.288, p < 0.01). Findings suggest that, in this context, affective activation (stimulation) may be a more proximal correlate of self-reported engagement than narrative immersion alone, warranting careful interpretation given the cross-sectional, self-report design. Full article

Background/Objectives: First-generation CDK4/6 inhibitors (palbociclib, ribociclib, abemaciclib) target the conserved ATP-binding pocket of CDK4 and, despite clinical success, are limited by acquired resistance and insufficient exploration of alternative regulatory sites. This study aimed to identify a putative allosteric small-molecule candidate at the CDK4 αE-helix–Cyclin D1 α1-helix protein–protein interaction (PPI) interface within the CDK4/Cyclin D1/p21 ternary complex using RapidFunnel-AI, a decision-interpretable virtual-screening pipeline. Methods: Starting from 50,000 ChEMBL 33 molecules, the pipeline sequentially applied a Q-Fold/RapidFunnel topological Tanimoto scan based on clinical CDK4/6 inhibitor motifs, fragment-level electronic-property enrichment, ADMET/PAINS filtering, dry Vina-GPU docking, hydration-mediated AutoDock-GPU (Version 1.6) docking, explicit-solvent molecular dynamics, contact-retention analysis, and MM-GBSA energy decomposition. The Q-Fold Thermo-Core surrogate model provided fragment-level enrichment, predicting the HOMO–LUMO gap (R² = 0.93) and isotropic polarizability (R² = 0.98) on QM9. Candidate selection did not rely on the lowest docking or MM-GBSA score alone, but on pose persistence, contact continuity, and energy-component consistency. Results: The workflow reduced the initial library to 43 topologically prioritized candidates, 25 ADMET/PAINS-filtered ligands, and 9 docking-derived complexes for MD validation. Ligand_020 emerged as the only candidate that preserved a persistent binding mode at Site 2 during a 500 ns simulation—an interface engagement reproduced across three independent 500 ns replicates with no full dissociation in any replicate—with a protein Cα RMSD of 2.88 ± 0.32 Å, a ligand heavy-atom RMSD of 3.56 ± 0.28 Å, and a van der Waals-dominated MM-GBSA profile (ΔGbind = −28.23 ± 3.57 kcal/mol). In contrast, palbociclib and ribociclib, forcibly placed at Site 2 as negative controls, lost most initial contacts within 5 ns and tended to detach despite more favorable MM-GBSA values. Conclusions: These results suggest that single-score docking or MM-GBSA ranking can generate false positives at shallow PPI interfaces. By integrating AI-assisted prioritization, multipocket docking, explicit-solvent MD, contact-retention analysis, and energy-component consistency, RapidFunnel-AI nominated Ligand_020 as an experimentally testable putative allosteric hit targeting the CDK4/Cyclin D1 interface, offering a reusable platform for PPI-focused oncological drug discovery. Full article

Background/Objectives: Fraxetin (7,8-dihydroxy-6-methoxycoumarin), a coumarin constituent of Cortex Fraxini (Qinpi) used in traditional Chinese medicine, is metabolised mainly by UGT1A9, but its potential to inhibit UGT enzymes and cause herb–drug interactions (HDIs) is largely unstudied. Methods: Fraxetin and four related coumarins were screened against 11 recombinant human UGTs; isoforms inhibited ≥80% underwent full kinetic analysis with 4-methylumbelliferone as probe. Binding was examined by molecular docking on AlphaFold structures with PLIP, triplicate 100 ns molecular dynamics, and MM/GBSA and MM/PBSA free-energy calculations, and interaction risk by FDA 2020 in vitro–in vivo extrapolation (IVIVE). Results: Fraxetin alone inhibited both UGT1A1 and UGT1A9 by >80% and was characterised in detail, acting as a mainly competitive mixed-type inhibitor (UGT1A1 IC₅₀ 15.99 μM, K_i 8.32 μM; UGT1A9 IC₅₀ 8.44 μM, K_i 5.90 μM). A structure–activity comparison identified a dual-element pharmacophore comprising the C-6 methoxy group and the 7,8-dihydroxycoumarin aglycone. MM/GBSA favoured UGT1A9 over UGT1A1 (ΔΔG = −4.06 kcal/mol, p = 0.005), concordant with the kinetic ranking. IVIVE predicted a borderline systemic signal (R₁ > 1.02) but an intestinal R_1,gut approximately five- to seven-fold above the high-risk threshold of 11 after capping the luminal concentration at fraxetin aqueous solubility. Conclusions: This is the first characterisation of fraxetin as a moderate-potency inhibitor of UGT1A1 and UGT1A9 and points to a previously under-recognised herb–drug interaction risk concentrated in the intestinal lumen rather than systemically; the finding constitutes an interaction signal requiring clinical confirmation rather than an established risk. Full article

Gelatin capsule waste (GCW), a protein-rich by-product, represents a promising substrate for the generation of potential bioactive substances, including free amino acids and other soluble substances generated during enzymatic hydrolysis. In this study, gelatin hydrolysates with degrees of hydrolysis (DH) ranging from 10% to 40% were produced using the commercial enzymes NS AC0106 (endopeptidase) and NS AC0107 (aminopeptidase) to enhance their functional properties. Increasing DH significantly improved antioxidant activity, surface hydrophobicity, and emulsifying capacity (p < 0.05), while sterilization further enhanced antioxidant capacity. Structural analyses confirmed extensive protein degradation and conformational modifications, as evidenced by SDS–PAGE (formation of low-molecular-weight substances), FTIR (shifts in the amide I region), and NMR (release of free amino acids). Electronic tongue analysis indicated that enzymatic hydrolysis enhanced umami and salty taste attributes. Notably, hydrolysis using NS AC0107 at 40% DH resulted in the highest antioxidant activity, together with pronounced umami taste and low bitterness. Overall, GCW-derived hydrolysates show considerable potential as functional ingredients and provide a sustainable strategy for the valorization of protein-rich industrial by-products. Full article

Robust age measurements for isolated neutron stars (NSs) are not easily available. That is why the characteristic age ${\tau }_{\mathrm{ch}}=P/2\dot{P}$ is often used as a proxy. Here, P is the spin period of the NS and $\dot{P}$ is the time derivative of P. Additional assumptions related to the initial properties and spin-down evolution are made to derive ${\tau }_{\mathrm{ch}}$. As a result, it is expected that ${\tau }_{\mathrm{ch}}$ is an upper limit for the real age ${\tau }_{\mathrm{real}}$. Recently, Chrimes et al. presented measurements of kinematic ages ${\tau }_{\mathrm{kin}}$ for several magnetars. Surprisingly, for the majority of these sources, ${\tau }_{\mathrm{kin}}>{\tau }_{\mathrm{ch}}$. We present a simple model that includes a realistic approximation for magnetic field decay in magnetars and a simple phenomenological description of field re-emergence following fallback after the birth of an NS. We demonstrate that this simple model can explain the observed relation ${\tau }_{\mathrm{kin}}>{\tau }_{\mathrm{ch}}$ for a realistic set of parameters. Full article

Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV), poses a serious threat to the global swine industry. Although modified live virus (MLV) vaccines have been widely used in the field for PRRS prevention for decades, the safety and efficacy of these vaccines have long been controversial. Here, we report a rare recombination pattern in China: the emergence of a novel NADC30-like PRRSV strain recombined from two MLV-like strains. Genome comparative analysis reveals that the SCMS2025 isolate has a non-continuous 136-amino acid deletion in the NSP2 protein and shares the highest nucleotide identity of 87.6% with lineage 5 (L5) strains. Phylogenetic analysis showed that SCMS2025 was classified into L1 (NADC30-like) strains based on ORF5 genotyping, whereas it belonged to a single branch between L1 and L5 strains based on the complete genomic sequences. Strikingly, genomic recombination analysis revealed that the newly emerged PRRSV isolate likely resulted from complex recombination events between NADC30-like and two MLV-like strains (RespPRRS MLV and TJbd14-1 MLV-like strains). Furthermore, SCMS2025 infection caused transient overt clinical signs followed by rapid recovery, indicating that the novel PRRSV isolate is a low pathogenic strain. Notably, all SCMS2025-inoculated piglets remained seronegative for PRRSV-specific antibodies throughout the entire 14-day observation period, suggesting a delayed onset of the host humoral immune response. Our study provides evidence for the ongoing evolution of PRRSV through inter lineage recombination and highlights the urgent need for safe and effective vaccines. Full article

Background: This study was conducted to identify genetic risk variants associated with nicotine addiction in the CHRNA5, HTR2A, DRD4, and CYP2A6 genes among electronic cigarette users who also smoke combustible cigarettes (dual users), and to assess potential genotoxic and cytotoxic damage in the oral mucosal cells of the study population. Methods: We included dual e-cig users (ECIG, n = 70), combustible cigarette smokers (CCU, n = 24), and non-smokers and non-e-cig users (NS, n = 110). Genetic variants in CHRNA5, HTR2A, DRD4, and CYP2A6 were genotyped. Micronucleus analysis was performed on oral mucosal cells to detect cellular abnormalities. Results: The ECIG group demonstrated greater nicotine addiction on the Fagerström Test for Nicotine Dependence (FTND, 5.5 vs. 1, p = 0.023). Salivary cotinine levels were significantly higher in the ECIG group compared to the CCU group (39 vs. 12 ng/mL, p < 0.001). The carriers of the A allele (rs16969968/CHRNA5) had higher FTND scores, carriers of the C allele (rs1800955/DRD4) used electronic cigarettes more frequently each day, and carriers of the T allele (rs4105144/CYP2A6) started using nicotine products at a younger age. The number of micronuclei and cellular abnormalities in the oral mucosa was higher in the ECIG and CCU groups compared to the NS group. Conclusions: Salivary cotinine levels and FTND are higher in dual e-cigarette users than in combustible cigarette users. Dual users exhibit risk alleles in the CHRNA5, DRD4, and CYP2A6 genes, which are associated with traits linked to increased nicotine addiction. Dual e-cigarette use poses comparable genotoxic risks to combustible smoking. Full article

Background: Ferrocene-containing compounds have gained attention in medicinal chemistry due to their unique redox and structural properties. This study investigates ferrocene-based analogues of imatinib and nilotinib to define their binding determinants within the ABL1 kinase domain using an integrated in silico approach, in relation to their previously reported cytotoxic activity. Methods: Ligand geometries were optimized at the B3LYP/def2-TZVP level with D3(BJ) dispersion and SMD solvation. Molecular docking against ABL1 (PDB ID: 2HYY) was performed using Glide SP, validated by re-docking and enrichment screening. Docked poses were refined using MM-GBSA (Prime, VSGB 2.1/OPLS4). The most active compounds (9 and 15a), together with the inactive control 15e, were subjected to three independent 500 ns molecular dynamics simulations (Desmond, OPLS4), followed by trajectory analysis including RMSD, RMSF, radius of gyration, SASA, and polar surface area. Results: Compounds 9 and 15a maintained stable binding within the ATP-binding pocket despite lacking the canonical hinge interaction with Met318, indicating hinge-independent binding. Their binding was mainly driven by interactions with Asp381 (DFG motif) and cation–π contacts with Lys271. In contrast, the compound 15e showed unstable binding, increased conformational flexibility, reduced pocket burial, and loss of key stabilizing interactions. Active compounds also preserved stable P-loop dynamics, with Tyr253 engagement suggesting a role in loop stabilization. Compound 9 exhibited the most constrained and reproducible binding mode among all analogues. Conclusions: Ferrocene-based analogues can sustain stable ABL1 binding via non-classical interaction networks independent of hinge recognition. The clear distinction between active compounds and the inactive analogue 15e supports the robustness of the proposed binding mode and provides a structural basis for their reported cytotoxic activity. These findings support further experimental evaluation of ferrocene-containing scaffolds as potential BCR-ABL1 inhibitors. Full article

O’nyong-nyong virus (ONNV) is a mosquito-borne alphavirus responsible for large-scale epidemics in sub-Saharan Africa. As the closest evolutionary relative of Chikungunya virus (CHIKV), ONNV shares substantial genetic similarity and overlapping clinical manifestations with CHIKV. Mechanistic understanding of ONNV infection has therefore largely been extrapolated from CHIKV rather than directly established. However, ONNV exhibits distinct biological features, including predominant transmission by Anopheles mosquitoes and a clinical presentation characterized by prominent lymphadenopathy with limited acute joint edema. These distinctions underscore the need for an integrated synthesis of experimentally validated determinants of ONNV infection. In this review, we summarize current evidence on molecular and immunological factors regulating ONNV infection in mammalian hosts and mosquito vectors. We first discuss species-specific viral clearance, host dependency factors, intrinsic antiviral restriction mechanisms, protective innate immunity, inflammatory pathology, and mechanism-informed therapeutic strategies in mammalian hosts. We then examine stage-specific immune regulation in Anopheles mosquitoes, emphasizing mechanisms that constrain viral replication while permitting persistent infection and transmission. Finally, we discuss nsP3-dependent vector specificity and the potential contribution of alternative mosquito species to ONNV ecology. Together, this review provides an integrated framework for understanding how host factors, immune responses, and vector-specific adaptations shape ONNV infection, pathogenesis, and transmission. Full article

Livestock intensification drives biodiversity loss, making impact quantification essential. Life Cycle Assessment (LCA) can evaluate whether regenerative practices, such as silvopastoral systems, mitigate this loss, but it requires specific characterization factors (CFs). In this pilot study, we applied the countryside Species-Area Relationship (SAR) model to derive the first invertebrate-specific CFs using dung beetles (Scarabaeinae). From field surveys, we calculated intensity-specific CFs for potential species loss (PSL/m²) in pastureland and cropland. We assessed biodiversity impacts per 1 kg calf live weight (LWC) across three livestock regimes: native silvopastoral (NSP, minimal land use), intensive silvopastoral (ISP, light land use), and monoculture (MC, intense land use). Results show high dung beetle affinity for NSP. The CFs distinguished impact intensity levels: MC had the highest PSL per area (6.76 × 10⁻¹⁰ PSL/m²), followed by ISP (5.93 × 10⁻¹⁰ PSL/m²) and NSP (4.99 × 10⁻¹⁰ PSL/m²). However, normalizing by yield reversed this trend: MC showed the lowest impact per 1 kg LWC (7.64 × 10⁻⁸ PSL/kg LWC), ISP was intermediate (1.06 × 10⁻⁷ PSL/kg LWC), and NSP had the highest impact (1.31 × 10⁻⁷ PSL/kg LWC). Incorporating upstream feed production significantly increased the overall biodiversity footprint, underscoring the need for comprehensive system boundaries. Integrating broader biodiversity components and landscape context remains essential to fully capture livestock management effects. Full article

Voltage-gated sodium channels are central to electrical excitability, and Nav1.7 is a major therapeutic target implicated in pain disorders and sensory signaling. Within the channel pore, permeating Na⁺ ions experience dynamically fluctuating hydration and coordination environments that may influence local ion–protein interactions. Identifying chemically distinct coordination states from molecular dynamics (MD) simulations is an important prerequisite for future higher-level electronic structure investigations. In this study, we present a reproducible workflow for identifying and classifying Na⁺ hydration–coordination microstates in the Nav1.7 pore using explicit-solvent molecular dynamics simulations. A geometrically defined pore region was used to quantify pore hydration and Na⁺ inner-shell coordination based on a 3.2 Å Na–O distance criterion. Na⁺ configurations were classified according to ligand identity into water-only (W), mixed protein–water (PW), and protein-only (P) microstates. Analysis of a 2 ns proof-of-principle simulation revealed a persistently hydrated pore environment, with Na⁺ coordination dominated by water-rich states and a smaller but distinct population of protein-contact configurations. These observations demonstrate that local coordination environments are chemically heterogeneous and cannot be fully described by hydration number alone. Representative structures from each microstate class were extracted to provide candidate configurations for future quantum mechanical, Quantum Mechanics/Molecular Mechanics (QM/MM), or density functional theory investigations of ion–ligand interactions in confined pore environments. The present work establishes a transparent and reproducible microstate-selection framework and does not report quantum mechanical energies, free-energy landscapes, or converged microstate populations. More broadly, the workflow provides a practical strategy for reducing complex MD ensembles into chemically interpretable coordination states suitable for subsequent higher-level analysis. Full article